Molecular Mechanism Of Fagopyri Dibotryis Rhizoma For The Treatment Of Cervical Cancer Based On Network Pharmacology And Molecular Docking

ObjectiveThe aim of this study was to use network pharmacology and molecular docking techniques,combined with in vitro experiments to initially validate the key targets and possible molecular mechanisms of cervical cancer treatment with Fagopyri dibotryis rhizoma,and to elucidate its mechanism of action.Methods1.Using network pharmacology techniques to analyse the potential biological pathways and key proteins of buckwheat for the treatment of cervical cancer;screening the potential active ingredients of buckwheat for the treatment of cervical cancer through molecular docking techniques and analyse the binding capacity of the active ingredients and key targets;using protein expression and purification techniques to obtain protein samples for the experiments in vitro;using surface plasmon resonance experiments to verify the binding capacity of the active ingredients and key proteins and binding mode.2.The effect of the active ingredient on the survival rate of tumour cells was observed by CCK8 assay;the effect of the active ingredient on gene expression in cervical cancer cells was determined by RT-PCR;the effect of the active ingredient on protein expression in cervical cancer cells was detected by Western Blot.Results1.Network pharmacology and molecular docking results: the use of network pharmacology techniques for Fagopyri dibotryis rhizoma;target proteins and cervical cancer target gene analysis found that tumor-related biological pathways play an important role in both;further comparative analysis of the biological pathways jointly regulated by the two,the results show that Extracellular signal-regulated kinase/mitogen-activated protein kinase(ERK/MAPK)pathway in Fagopyri dibotryis rhizoma;and a visual analysis of the ERK/MAPK pathway revealed that the central region of the network was composed of Mitogen-activated protein kinase 1(MAPK1),Signal transducers and activators of transcription 3(STAT3),and a number of other pathways.MAPK1,STAT3,estrogen receptor 1(ESR1),C-RAF proto-oncogene serotonin kinase(RAF1),and SRC protein kinase(SRC)are key targets that act together,and MAPK1 and other proteins with the highest degree of connectivity;the screening of the active ingredients of Fagopyri dibotryis rhizoma;and key targets for molecular docking found that catechin gallate and MAPK1 binding energy lowest affinity for the strongest,which may be the potential active ingredient of Fagopyri dibotryis rhizoma;for cervical cancer;catechin gallate and cervical cancer crossover targets for network pharmacology analysis found that the ERK/MAPK biological pathway The ERK/MAPK pathway was the most strongly associated with both;the binding ability of catechin gallate to MAPK1 was verified by surface plasmon resonance experiments,and specific binding between the two was found to exist,and the binding ability was strong.2.In vitro results: The effects of different concentrations of catechin gallate on cervical cancer Hela cells were investigated,and the results showed that catechin gallate had a significant inhibitory effect on the survival rate of cervical cancer Hela cells,and the cell survival rate tended to decrease with the increase of drug concentration,with an IC50 of 100 μM;for Hela cells intervened with catechin gallate,compared with For Hela cells treated with catechin gallate,Tumour Necrosis Factor α(TNF-α),B-cell lymphoma/leukemia 2(Bcl-2)m RNA expression was down-regulated in the 50 μM and100 μM catechin gallate intervention groups compared to the Control group,while the pro The m RNA expression of BCL2-Associated X(Bax)was up-regulated;at the same time,50 μM and 100 μM catechin gallate intervention groups could inhibit the phosphorylation levels of MAPK1,Inhibitor of NF-κB kinase α(IKKα),Inhibitor of NF-κb α(IκBα),Nuclear factor-κB(NF-κB),thereby blocking activation of the ERK/MAPK pathway.ConclusionThe active ingredient in Fagopyri dibotryis rhizoma,catechin gallate,may affect the ERK/MAPK pathway by targeting MAPK1,which may be one of the potential mechanisms for cervical cancer treatment with buckwheat.